×

NONINVASIVE DIAGNOSTICS BY SEQUENCING 5-HYDROXYMETHYLATED CELL-FREE DNA

  • US 20190017109A1
  • Filed: 09/28/2018
  • Published: 01/17/2019
  • Est. Priority Date: 04/07/2016
  • Status: Active Grant
First Claim
Patent Images

1. A method for sequencing hydroxymethylated cell-free DNA (cfDNA),(a) adding adaptor sequences onto the ends of cfDNA;

  • (b) incubating the adaptor-ligated cfDNA with a DNA β

    -glucosyltransferase and UDP glucose modified with a chemoselective group, thereby covalently labeling the hydroxymethylated DNA molecules in the cfDNA with the chemoselectivegroup;

    (c) linking a biotin moiety to the chemoselectively-modified cfDNA via a cycloaddition reaction;

    (d) enriching for the biotinylated DNA molecules by binding the product of step (c) to a support that binds to biotin;

    (e) amplifying the enriched DNA using primers that bind to the adaptors; and

    (f) sequencing the amplified DNA to produce a plurality of sequence reads, wherein the method does not comprise releasing the biotinylated DNA molecules from the support after step (d), prior to step (e).

View all claims
  • 3 Assignments
Timeline View
Assignment View
    ×
    ×